Effects of topical ketorolac tromethamine on tear parameters, meibography, goblet cell density, and conjunctival oxidative stress in healthy dogs.

OBJECTIVES: The objective of the study was to evaluate whether a twice-daily instillation of 0.45% preservative-free ketorolac tromethamine (FKT) or 0.4% benzalkonium chloride-preserved ketorolac tromethamine (BACKT), every 12 h for 30 days may affect tear film parameters and the meibography in healthy dogs. Additionally, we assessed whether the same treatments irritated the ocular surface, affected goblet cell density (GCD), and the levels of oxidative stress biomarkers (OSB) in the conjunctiva of the same dogs. PROCEDURES: Experimental and masked comparison study. In 11 healthy dogs baseline values of the lipid layer thickness, tear meniscus height, non-invasive tear breakup time (NI-TFBT), and the meibomian gland (MG) loss were assessed by OSAvet®. For each dog, one eye received 40 µL of BACKT, while the other received 40 µL FKT, every 12 h for 30 consecutive days. Tear parameters and meibography were repeated 15, 30, and 60 days post-treatments. Conjunctival hyperemia and blepharospasm were monitored at the same time points. At baseline and Day 30, a conjunctival biopsy was collected for GCD and OSB determination. RESULTS: Conjunctival hyperemia and blepharospasm were not observed. At Day 15, the MG loss increased only in FKT-treated eyes (p < .001). On Day 30, both treatment groups showed increased MG loss, shortened NI-TFBT, and reduced GCD and catalase (p < .05). At Day 30, BACKT-treated eyes showed lower levels of superoxide dismutase (SOD) (p = .006) and higher levels of malondialdehyde (MDA) (p = .02). Differences between treatments were not observed for any parameter at any time point (p > .05). 60 days after treatment, OSAvet® parameters tended to return to values assessed at baseline; however, significant differences remained for MG loss (p < .05). CONCLUSIONS: Twice-daily instillation of KT, containing or not BAC, for 30 consecutive days shortened NI-TFBT, decreased GCD, and increased the MG loss in healthy dogs. KT should be used with caution when prescribed for long periods, particularly in patients with tear film abnormalities. However, future controlled studies using KT, BAC, and other topical NSAIDs are indicated to further support this finding.

Effects of commercial amniotic membrane extract on the re-epithelialization time and the early expression of matrix metalloproteinase-9 in cats with experimentally induced corneal ulcers.

OBJECTIVES: To investigate whether a commercially available amniotic membrane extract (AME) can accelerate corneal wound healing and suppress the early expression of MMP-9 in the tears of cats with experimentally induced superficial ulcerative keratitis. PROCEDURES: A total number of 16 cats were included. At the end of keratectomy, cats in the treatment group (TG, n = 8) received 40 µl of AME (EyeQ® Amniotic Eye Drops, Vetrix®) four times daily, while cats in the control group (CG, n = 8) received 40 µl of saline at the same time points. Tears were collected 24 and 48 h after keratectomy, and the total MMP-9 was quantified by ELISA. RESULTS: The corneal re-epithelialization rate did not differ between groups (p = .26), being 0.48 ± 0.05 mm2 /h in the CG and 0.41 ± 0.03 mm2 /h in the TG. Similarly, the average time to achieve corneal wound healing did not differ between groups (p = .25) and was 61.50 ± 3.54 h in the CG and 70.50 ± 6.71 h in the TG. The dimensions of the ulcerated areas also did not differ at any time point between the groups (p > .05). In both groups, corneas healed without scarring, pigmentation, or vascularization. The expression of MMP-9 in the tears was similar in both groups at 24 h post-keratectomy, with a slight decrease at 48 h (p > .05). CONCLUSIONS: The instillation of a commercial AME (EyeQ®) is safe, but it did not decrease the corneal re-epithelialization time or the early expression of MMP-9 in the tears of cats with experimentally induced superficial ulcerative keratitis in this study.

Effect of 0.15% sodium hyaluronate on tear film breakup time in healthy anesthetized cats.

OBJECTIVES: This study aimed to evaluate the effect of 0.15% sodium hyaluronate (SH) on tear film breakup time (TFBT) in healthy anesthetized cats. PROCEDURES: Forty cats undergoing elective surgery were enrolled. TFBT was assessed before anesthesia to obtain baseline values. As a preanesthetic medication, cats received midazolam, tramadol, and cetamine combined in the same syringe. For anesthetic induction and maintenance, propofol and isoflurane were used. After a 15 min stabilization period to achieve the surgical anesthetic plane, one eye was treated with one drop of SH, while the other eye received saline and served as a control. TFBT was measured at the end of the general anesthesia (T40) and 35 (T75) and 80 min (T120) after the termination of the anesthesia. TFBT values were compared between the control and SH-treated eyes; both values were also compared with the baseline values (p < .05). RESULTS: In the control eyes, TFBT significantly decreased from baseline at all time points (p < .001), while in SH-treated eyes, TFBT significantly increased from baseline only at T40 (p < .0001). In SH-treated eyes, TFBT was significantly higher than that in the control eyes at all time points (p < .001). CONCLUSIONS: In healthy cats, TFBT decreases significantly after 40 min of general anesthesia, and one drop of 0.15% sodium hyaluronate was able to maintain the stability of the tear film for up to 75 min in treated eyes. However, the isolated effect of each drug used in our anesthetic protocol on TFBT should be executed in further studies.

Concentrations of tissue inhibitor of matrix metalloproteinase-1 and hyaluronic acid in canine amniotic membranes cryopreserved for different time points and its effects in dogs with complicated corneal ulcers.

OBJECTIVES: To determine the concentrations of total protein (TP), tissue inhibitor of matrix metalloproteinase-1 (TIMP-1) and hyaluronic acid (HA) in amniotic membranes (AMs) harvested from placentas of bitches of different ages and cryopreserved for different time points. The outcomes of complicated corneal defects of dogs repaired with AMs stored for the same time points were also evaluated. PROCEDURES: Ten cryopreserved canine AMs were stored for short term (2-50 days), middle term (92-210 days), or long term (256-357 days). TP was quantified by Bradford's test, whereas TIMP-1 and HA were quantified by ELISA. Twenty-one dogs that had an AM transplantation to restore deep or perforating corneal wounds were selected. RESULTS: TIMP-1 levels were lower in AMs cryopreserved for middle term (p = .02) and long term (p = .0009), when compared to AMs stored for short term. TP (p = .39) and HA (p = .18) concentrations in AMs did not differ among the storage time. TIMP-1 concentration in AMs correlated with storage time (R = -.65, p < .0001), while TP (R = -.33, p = .07) and HA concentrations did not (R = -.15, p = .41). The age of donors did not correlate with the components evaluated in the AMs. Corneal defects repaired with AMs stored for short term healed sooner than the ones repaired with AMs stored for middle (p < .01) and long term (p = .02). Additionally, TIMP-1 levels in AMs correlated negatively with the epithelization time (R = -.62, p = .002). Graft opacity was severe in 55% of cases. However, the HA levels in AMs correlated negatively with the opacification score (R = -.47, p = .03). Vision was observed in more patients who presented deep ulcers and descemetoceles, than in the ones with perforations (p = .004). CONCLUSIONS: TIMP-1 concentration in canine AMs significantly decreased over a year storage time, while TP and HA concentrations did not change during the same period. The age of donors did not correlate with the components evaluated in the AMs. Complicated corneal defects repaired with AMs cryopreserved for short term healed sooner and tended to be less opaque; however, satisfactory to optimal outcomes were achieved even in the eyes repaired with AMs stored for up to a year.

Evaluation of barbed suture for celiorrhaphy and subcutaneous closure in bitches with pyometra submitted to ovariohysterectomy.

PURPOSE: To evaluate the use of barbed sutures over the surgical time, the leukogram, the tissue thickness in which the sutures were employed (ultrasonography), the costs, and the possible complications in bitches with pyometra submitted to ovariohysterectomy (OH). METHODS: Convectional 2.0 polyglyconate suture was used in the control group (CG n = 10) and 2.0 barbed polyglyconate suture in the barbed group (BG n = 10) to perform celiorrhaphy (simple continuous pattern) and subcutaneous closure (continuous intradermal pattern). Data were assessed using paired (leukogram between 24 and 48 h within the same group) and unpaired (leukogram, surgical time, tissue thickness, and costs) Student's t-test. The Fisher exact test was used to assess the occurrence of seroma between groups (p < 0.05). Results are shown as mean ± standard error of mean. RESULTS: The time spent to perform the celiorrhaphy (195.30 ± 17.37 s vs. 204 ± 16.00 s), subcutaneous closure (174.0 ± 15.86 s vs. 198.0 ± 15.62 s), and the total surgical time (24.30 ± 1.44 min vs. 23.00 ± 1.30 min) did not differ between BG and CG, respectively (p > 0.05). Leukogram at 48 h post-surgery did not differ between groups (p = 0.20). No differences were observed in the subcutaneous and the abdominal wall thickness (cm) assessed by ultrasonography at 48 h in BG (0.31 ± 0.04, 0.80 ± 0.05) and CG (0.34 ± 0.03, 0.72 ± 0.06), respectively. Similarly, 15 days post-surgery the same structures did not differ between BG (0.26 ± 0.02, 0.74 ± 0.08) and CG (0.26 ± 0.03, 0.64 ± 0.05) (p > 0.05). In one bitch from each group, a mild seroma was observed on one side of the surgical wound 48 h after surgery (p = 1.00). The procedures in which barbed sutures were used had an average additional cost of R$ 200.00 ± 11.66 (p < 0.0001). CONCLUSIONS: Barbed suture has proven to be efficient and safe for abdominal and subcutaneous closure. However, considering its current high cost in addition thatthe surgical time of bitches with pyometra undergone OH was not reduced, no advantages were observed with theuse of barbed sutures for this type of surgery.

Evaluation of barbed suture for celiorrhaphy and subcutaneous closure in bitches with pyometra submitted to ovariohysterectomy

ABSTRACT Purpose To evaluate the use of barbed sutures over the surgical time, the leukogram, the tissue thickness in which the sutures were employed (ultrasonography), the costs, and the possible complications in bitches with pyometra submitted to ovariohysterectomy (OH). Methods Convectional 2.0 polyglyconate suture was used in the control group (CG n = 10) and 2.0 barbed polyglyconate suture in the barbed group (BG n = 10) to perform celiorrhaphy (simple continuous pattern) and subcutaneous closure (continuous intradermal pattern). Data were assessed using paired (leukogram between 24 and 48 h within the same group) and unpaired (leukogram, surgical time, tissue thickness, and costs) Student's t-test. The Fisher exact test was used to assess the occurrence of seroma between groups (p < 0.05). Results are shown as mean ± standard error of mean. Results The time spent to perform the celiorrhaphy (195.30 ± 17.37 s vs. 204 ± 16.00 s), subcutaneous closure (174.0 ± 15.86 s vs. 198.0 ± 15.62 s), and the total surgical time (24.30 ± 1.44 min vs. 23.00 ± 1.30 min) did not differ between BG and CG, respectively (p > 0.05). Leukogram at 48 h post-surgery did not differ between groups (p = 0.20). No differences were observed in the subcutaneous and the abdominal wall thickness (cm) assessed by ultrasonography at 48 h in BG (0.31 ± 0.04, 0.80 ± 0.05) and CG (0.34 ± 0.03, 0.72 ± 0.06), respectively. Similarly, 15 days post-surgery the same structures did not differ between BG (0.26 ± 0.02, 0.74 ± 0.08) and CG (0.26 ± 0.03, 0.64 ± 0.05) (p > 0.05). In one bitch from each group, a mild seroma was observed on one side of the surgical wound 48 h after surgery (p = 1.00). The procedures in which barbed sutures were used had an average additional cost of R$ 200.00 ± 11.66 (p < 0.0001). Conclusions Barbed suture has proven to be efficient and safe for abdominal and subcutaneous closure. However, considering its current high cost in addition thatthe surgical time of bitches with pyometra undergone OH was not reduced, no advantages were observed with theuse of barbed sutures for this type of surgery.

Effects of intracameral injection of epinephrine and 2% lidocaine on pupil diameter, intraocular pressure, and cardiovascular parameters in healthy cats.

OBJECTIVES: To investigate the effects of the intracameral injection of epinephrine and two doses of 2% lidocaine on pupil diameter (PD), intraocular pressure (IOP), heart rate (HR), and mean arterial pressure (MAP) in healthy cats. METHODS: Five treatment groups were formed (10 cats/each). Animals received 0.2 mL of epinephrine, 0.2 or 0.3 mL of 2% lidocaine, or 0.2 mL of BSS. Cats were anesthetized, and all solutions were injected intracamerally. PD, IOP, HR, and MAP were assessed at baseline, following anterior chamber paracentesis (T0), and at every 5 minutes, until anesthesia was terminated (T60). PD and IOP continued to be assessed for 2 additional hours during recovery from anesthesia. In another group, cats were not anesthetized and one of the eyes was treated with one drop of 0.5% tropicamide to check for maximal pupil diameter. RESULTS: Faster onset and longer duration of sufficient mydriasis (>10 mm) were observed in epinephrine treatment group, when comparing with cats treated with both doses of lidocaine. Eyes treated with epinephrine achieved the largest maximum pupil diameter (mm) when comparing with eyes treated with 0.3 mL of lidocaine (11.01 ± 0.16), tropicamide (10.66 ± 0.17), and 0.2 mL of lidocaine (10.23 ± 0.12) (P < .0001). In all groups, IOP decreased significantly at T0, but tended to return to baseline at T60. HR and MAP did not change significantly during time and among treatments. CONCLUSIONS: The intracameral injection of 0.2 mL of 1:100 000 epinephrine and 0.3 mL of 2% lidocaine can be used as an alternative to tropicamide in healthy cats. Both treatments produced satisfactory and long-lasting mydriasis without adverse effects on IOP, HR, and MAP.

Effects of tramadol and acepromazine on intraocular pressure and pupil diameter in young healthy cats / Efeitos do tramadol e da acepromazina sobre a pressão intraocular e o diâmetro pupilar em gatos saudáveis jovens

ABSTRACT: This study aimed to investigate the effects of the systemic administration of acepromazine, tramadol and the association of both on intraocular pressure (IOP) and pupil diameter (PD) in young healthy cats. Cats were randomly allocated into three groups (n=10/each) and intramuscular acepromazine (AG), tramadol (TG) or acepromazine combined with tramadol (ATG) were injected. PD (electronic caliper) and IOP (applanation tonometry) were assessed before (baseline) and following 15, 30, 60, and 120 minutes of treatments. It was verified that in AG, PD decreased significantly from time point 30 to 120 (P=0.002), but such reduction did not differ significantly from baseline (P=0.89). In TG, PD increased significantly from the first 15 minutes, until the last time point of evaluation (P<0.001). In ATG, PD increased significantly from time point 30 to 120 when compared to baseline (P<0.001); but significant differences from time point 30 to 120 were not seen (P=0.71). Comparisons among groups showed that PD values of TG and ATG were significantly higher than that of AG (P<0.05). IOP values, on the other hand, did not change significantly among time points and groups (P>0.05). It can be concluded that tramadol alone or in association with acepromazine produced significant mydriasis for up to 120 minutes, without changing IOP values in normal cats. Results of this study suggested that tramadol alone or in association with acepromazine caused significant mydriasis and did not change IOP values in normal cats. Therefore, it may be considered a satisfactory pre-anesthetic combination for ophthalmic surgery in cats. However, further studies are warranted on the use of such protocols in cats with ophthalmic diseases undergoing ocular or intraocular surgery.

RESUMO: Objetivou-se estudar os efeitos da administração sistêmica da acepromazina, do tramadol e da associação de ambos sobre a pressão intraocular (PIO) e o diâmetro pupilar (DP) em gatos saudáveis jovens. Os gatos foram aleatoriamente distribuídos em três grupos (n=10/cada) e tratados pela via intramuscular com acepromazina (GA), tramadol (GT) ou acepromazina combinada ao tramadol (GAT). O DP (paquimetria eletrônica) e a PIO (tonometria de aplanação) foram mensurados antes (basal) e após 15, 30, 60, e 120 minutos após a administração dos tratamentos. Constatou-se que no GA, o DP reduziu significativamente a partir do 30˚ até o 120˚ minuto de avaliação (P=0.02), mas sem diferir significativamente em relação ao basal (P=0,89). No GT, o DP se elevou significativamente desde 15˚ minuto, até o último período de avaliação (P˂0.001). No GAT, o DP se elevou de forma significativa do 30˚ ao 120˚ minuto em comparação ao basal (P<0,001), mas esse parâmetro não alterou significativamente do 30º ao 120º minuto (P=0.71). Comparações entre os grupos mostraram que o DP do GT e do GAT apresentaram valores significativamente mais elevados que aqueles do GA (P<0,05). A PIO, por sua vez, não se alterou de forma significativa nos períodos e entre os grupos avaliados (P>0,05). Conclui-se que o tramadol, administrado de forma isolada ou em associação à acepromazina, produz midríase de até 120 minutos, sem alterar os valores da PIO em gatos saudáveis. Dessa forma, esse protocolo pré-anestésico pode ser considerado uma alternativa para cirurgia oftálmica em gatos. Todavia, os resultados desse protocolo em gatos com doença oftálmica, que necessitem de cirurgia, devem ser avaliados em estudos futuros.

Effects of dorzolamide/timolol and tafluprost on intraocular pressure and pupil diameter in healthy dogs / Efeitos da dorzolamida/timolol e da tafluprosta sobre a pressão intraocular e o diâmetro pupilar em cães saudáveis

ABSTRACT: This study aimed to evaluate and compare the effects of the fixed combination of dorzolamide/timolol with those of tafluprost on intraocular pressure (IOP) and pupil diameter (PD) in healthy dogs (n=10). Two experiments were conducted with an interval of 30 days. In both, IOP and PD were assessed at 8, 11, 14, 17, and 20h. Parameters were evaluated during baseline, treatment period of four days, and one day of post-treatment. During treatment phase, IOP decreased by 0.74 (P<0.05), 1.88 (P<0.01), 2.94 (P<0.001), and 3.10mmHg (P<0.01), in dorzolamide/timolol-treated eyes; and by 1.50, 2.18, 2.14, and 2.18mmHg (P<0.001), in tafluprost-treated eyes. PD decreased by 0.24 (P<0.01), 0.32 (P<0.01), 0.49 (P<0.001), and 0.40mm (P<0.001), in dorzolamide/timolol treated eyes; and by 2.31, 2.55, 2.43, and 2.70mm (P<0.001), in tafluprost-treated eyes. Dorzolamide/timolol and tafluprost were able to decrease IOP and PD in healthy dogs. However, a cumulative effect of the fixed combination of dorzolamide/timolol was more effective in reducing IOP, than tafluprost. Comparisons between treatments showed that tafluprost was more effective in reducing PD throughout the treatment phase.

RESUMO: O estudo objetivou avaliar e comparar os efeitos da combinação fixa da dorzolamida/timolol com os da tafluprosta sobre a pressão intraocular (PIO) e o diâmetro pupilar (DP) em cães saudáveis (n=10). Dois experimentos com intervalo de 30 dias foram conduzidos. Em ambos, a PIO e o DP foram avaliados às 8, 11, 14, 17 e às 20h. Os parâmetros foram avaliados durante a fases basal, um período de tratamento de quatro dias, e um dia de pós-tratamento. Durante a fase de tratamento, a PIO dos olhos tratados com dorzolamida/timolol reduziram em 0.74 (P<0.05), 1.88 (P<0.01), 2.94 (P<0.001), e 3.10mmHg (P<0.01); e dos olhos tratados com tafluprosta em 1.50, 2.18, 2.14 e 2.18mmHg (P<0.001). O DP dos olhos tratados com dorzolamida/timolol reduziram em 0.24 (P<0.01), 0.32 (P<0.01), 0.49 (P<0.001) e 0.40mm (P<0.001); e dos olhos tratados com tafluprosta em 2.31, 2.55, 2.43 e 2.70mm (P<0.001). A dorzolamida/timol e a tafluprosta foram capazes de reduzir a PIO e o DP em cães saudáveis. Porém, efeito cumulativo do tratamento com dorzolamida/timolol foi observado, decorridos três dias de tratamento. Por essa razão, a dorzolamida/timolol foi mais efetiva que a tafluprosta na redução da PIO. Comparações entre os tratamentos demonstraram que a tafluprosta foi mais efetiva em reduzir o DP, durante toda a fase de tratamento.

Firocoxib on aqueous humor prostaglandin E 2 levels for controlling experimentally-induced breakdown of blood-aqueous barrier in healthy and Toxoplasma gondii -seropositive cats / Efeitos do firocoxib sobre os níveis de prostaglandina E 2 no humor aquoso de gatos saudáveis e com sorologia positiva para toxoplasmose mediante quebra da barreira hematoaquosa experimentalmente induzida

ABSTRACT: This study aimed to evaluate the effects of firocoxib for controlling experimentally-induced breakdown of the blood-aqueous barrier in healthy and Toxoplasma gondii -seropositive cats. Thirty two cats with no ocular abnormalities were used. Groups (n=8/each) were formed with healthy cats that received 5mg g-1 of oral firocoxib (FH) or no treatment (CH) on day 0; seropositive cats for anti -T. gondii specific immunoglobulin G (IgG) were grouped (n=8/each) and treated in a similar fashion (FT and CT). On day 1, cats of all groups received the same treatment protocol, and 1h later, aqueocentesis was performed under general anesthesia (M0). Following 1h, the same procedure was repeated (M1). Quantitation of aqueous humor total protein and prostaglandin E2 (PGE2) were determined. Aqueous samples of seropositive cats were tested for anti- T. gondii specific IgG. In M0, aqueous samples of CT showed a significantly higher concentration of PGE2 in comparison with other groups (P<0.05). In all groups, PGE2 concentration increased significantly from M0 to M1 (P=0.001). PGE2 values did not change significantly between groups in M1 (P=0.17). Anti- T. gondii specific IgG were reported only in samples of M1, and aqueous titers did not change significantly between FT and CT (P=0.11). Although we have observed that aqueous humor PGE2 levels were significantly higher in cats of CT group during M0, such increase was not able to break the blood-aqueous barrier and cause anterior uveitis. Firocoxib did not prevent intraocular inflammation after aqueocentesis, in healthy and toxoplasmosis-seropositive cats.

RESUMO: Objetivou-se avaliar a eficácia do firocoxib no controle da quebra da barreira hematoaquosa experimentalmente induzida em gatos saudáveis e com sorologia positiva para toxoplasmose. Para tanto, utilizaram-se trinta e dois gatos sem alterações oculares, alocados em grupos (n=8/cada) compostos por gatos saudáveis que receberam tratamento prévio com 5mg g-1 de firocoxib oral (HF) ou sem nenhum tratamento (CH) no dia 0, e por gatos com sorologia positiva para toxoplasmose tratados de maneira similar (FT e CT). No dia 1, os gatos de todos os grupos receberam o mesmo protocolo de tratamento do dia anterior e, 1h depois, foram submetidos à paracentese da câmara anterior sob anestesia geral (M0). Após 1h, realizou-se nova paracentese (M1). Mediante a colheita de humor aquoso (M0 e M1), quantificaram-se os valores de proteína total e prostaglandina E2 (PGE2) das amostras. As amostras dos gatos com sorologia positiva para toxoplasmose foram também testadas para anticorpos anti- T. gondii IgG específicos. Em M0, as amostras de humor aquoso de CT apresentaram concentração de PGE2 significativamente superior aos demais grupos (P<0,05). Em todos os grupos, a concentração de PGE2 aumentou significativamente de M0 para M1 (P=0,001), no entanto, não houve diferença significativa entre os grupos em M1 (P=0,17). Anticorpos anti -T. gondii IgG específicos foram encontrados somente em amostras de M1, e os títulos não diferiram significativamente entre FT e CT (P=0,11). Valores de PGE2 significativamente superiores no CT durante M0 não foram capazes de induzir a quebra da barreira hematoaquosa e causar uveíte anterior nos gatos deste estudo. O firocoxib, por sua vez, não foi capaz de prevenir a quebra da barreira hematoaquosa após realização de paracente na câmara anterior em gatos saudáveis e com sorologia positiva para toxoplasmose.

Effects of morphine on the expression of cytokines and inflammatory mediators in a rabbit model of endotoxin-induced experimental uveitis.

PURPOSE: To evaluate the effects of 1% morphine instillation on clinical parameters, aqueous humor turbidity, and expression levels of tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1beta), prostaglandin E2 (PGE2), and myeloperoxidase (MPO) in rabbits with endotoxin-induced experimental uveitis. METHODS: Twenty four New Zealand white rabbits were divided into four groups (n=6 each): control (CG), morphine (MG), naloxone (NG), and morphine-naloxone (MNG) groups. Under dissociative anesthesia, 0.1 mL of solution containing 0.2 µg of lipopolysaccharide (LPS) endotoxin from the Salmonella typhimurium cell wall was injected in the vitreous chamber. Clinical evaluations (conjunctical hyperemia, chemosis blepharospasm, and ocular discharge) and laser flaremetry were performed before (baseline), and 10 and 20 hours after induction of uveitis. Rabbits were subsequently euthanized and eyes were enucleated to quantify expression levels of TNF-α, IL-1 beta, PGE2, and MPO. RESULTS: No significant differences in clinical parameters and flare values were observed between the study groups. TNF-α and IL-1 beta levels increased significantly in the CG, MG, NG, and MNG groups compared to baseline (P<0.05). Significant differences in PGE2 levels were observed between the MG and NMG groups (P<0.05). A trend toward increased MPO activity was observed in response to uveitis induction; however, this trend did not reach statistical significance (P>0.05). CONCLUSIONS: Morphine has no effect on clinical parameters, flare, or expression levels of inflammatory mediators in a rabbit model of uveitis induced by intravitreal injection of LPS.

Effects of morphine on the expression of cytokines and inflammatory mediators in a rabbit model of endotoxin-induced experimental uveitis / Efeitos da morfina na expressão de citocinas e de mediadores inflamatórios em coelhos com uveítes experimental induzida por endotoxina

ABSTRACT Purpose: To evaluate the effects of 1% morphine instillation on clinical parameters, aqueous humor turbidity, and expression levels of tumor necrosis factor alpha (TNF-α), interleukin-1 beta (IL-1beta), prostaglandin E2 (PGE2), and myeloperoxidase (MPO) in rabbits with endotoxin-induced experimental uveitis. Methods: Twenty four New Zealand white rabbits were divided into four groups (n=6 each): control (CG), morphine (MG), naloxone (NG), and morphine-naloxone (MNG) groups. Under dissociative anesthesia, 0.1 mL of solution containing 0.2 µg of lipopolysaccharide (LPS) endotoxin from the Salmonella typhimurium cell wall was injected in the vitreous chamber. Clinical evaluations (conjunctical hyperemia, chemosis blepharospasm, and ocular discharge) and laser flaremetry were performed before (baseline), and 10 and 20 hours after induction of uveitis. Rabbits were subsequently euthanized and eyes were enucleated to quantify expression levels of TNF-α, IL-1 beta, PGE2, and MPO. Results: No significant differences in clinical parameters and flare values were observed between the study groups. TNF-α and IL-1 beta levels increased significantly in the CG, MG, NG, and MNG groups compared to baseline (P<0.05). Significant differences in PGE2 levels were observed between the MG and NMG groups (P<0.05). A trend toward increased MPO activity was observed in response to uveitis induction; however, this trend did not reach statistical significance (P>0.05). Conclusions: Morphine has no effect on clinical parameters, flare, or expression levels of inflammatory mediators in a rabbit model of uveitis induced by intravitreal injection of LPS.

RESUMO Objetivo: Estudaram-se os efeitos da instilação de morfina 1% sobre parâmetros clínicos, turbidez do humor aquoso e expressão de fator de necrose tumoral alfa (TNF-alfa), de interleucina-1 beta (IL-1beta), de prostaglandina E2 (PGE2) e de mieloperoxidase (MPO), em olhos de coelhos com uveíte induzida por endotoxina. Material e Métodos: Vinte e quatro coelhos da raça Nova Zelândia Branco foram distribuídos em quatro grupos (n=6, em cada): grupo controle (GC), morfina (GM), naloxona (GN) e morfina-naloxona (GMN). Sob anestesia dissociativa, injetou-se 0,1 mL de solução contendo 0,2 µg de lipossacarídeo (LPS) endotóxico da parede celular de Salmonella typhimurium na câmara vítrea. Realizou-se avaliação clínica (hiperemia conjuntival, quemose, blefaroespasmo e secreção ocular) e a flaremetria a “laser” antes (basal) e após 10 e 20 horas da indução da uveíte. No final, os coelhos foram submetidos à eutanásia e os olhos com uveíte foram enucleados para a quantificação dos níveis de TNF-alfa, IL-1 beta, PGE2 e MPO. Diferenças foram consideradas significativas quando p<0,05. Resultados: Os grupos da pesquisa não diferiram quanto aos parâmetros clínicos e os valores de “flare”. Observou-se elevação significativa nos níveis de TNF-alfa e de IL-1 beta, comparativamente ao basal, nos grupos GC, GM, GN e GMN (p<0,05). Valores de PGE2 variaram entre os grupos GM e GNM (p<0,05). A atividade de MPO aumentou após a indução da uveíte, porém, sem significância estatística (p>0,05). Conclusões: A morfina não atuou sobre parâmetros clínicos, “flare” e expressão dos mediadores inflamatórios estudados, quando instilada em olhos de coelhos com uveíte induzida por injeção intravítrea de LPS.

Evaluation of intraluminal pressure in cystorrhaphies with and without intestinal serosal patch supplementation from canine cadavers.

PURPOSE: To evaluate the maximal intraluminal pressure (MIP) supported by canine cadaveric urinary bladders that underwent cystotomy followed by cystorraphy, with and without serosal patching-supplementation. METHODS: Two groups (n=8 each) were formed, and in one (conventional) the cystotomy was closed with cushing pattern. In the other group (serosal), the same procedure was performed, and a piece of jejunum was used for the construction of the serosal patching over the cystorraphy. MIP was measured by means of an invasive blood pressure transducer with closed stopcock attached to a multiparameter monitor. At the end of each measurement, the bladder body circumference was assessed. RESULTS: Mean ± SD MIP sustained for the conventional and serosal groups were 28.88 ± 5.08 and 65.38 ± 10.99 mmHg, respectively (p < 0.0001). Bladder circumference did not change significantly between groups (p = 0.35) and did not correlate with MIP assessed in conventional (p = 0.27; r = 0.4379) and serosal groups (p = 0.37; r = -0.3637). CONCLUSION: Serosal patch-supplemented cystorraphies were able to sustain intraluminal pressures 55.8% higher, than nonsupplemented cystorraphies in specimens from canine cadavers.

Evaluation of intraluminal pressure in cystorrhaphies with and without intestinal serosal patch supplementation from canine cadavers

PURPOSE:To evaluate the maximal intraluminal pressure (MIP) supported by canine cadaveric urinary bladders that underwent cystotomy followed by cystorraphy, with and without serosal patching-supplementation.METHODS:Two groups (n=8 each) were formed, and in one (conventional) the cystotomy was closed with cushing pattern. In the other group (serosal), the same procedure was performed, and a piece of jejunum was used for the construction of the serosal patching over the cystorraphy. MIP was measured by means of an invasive blood pressure transducer with closed stopcock attached to a multiparameter monitor. At the end of each measurement, the bladder body circumference was assessed.RESULTS:Mean±SD MIP sustained for the conventional and serosal groups were 28.88±5.08 and 65.38±10.99 mmHg, respectively (p<0.0001). Bladder circumference did not change significantly between groups (p=0.35) and did not correlate with MIP assessed in conventional (p=0.27; r=0.4379) and serosal groups (p=0.37; r=-0.3637).CONCLUSION:Serosal patch-supplemented cystorraphies were able to sustain intraluminal pressures 55.8% higher, than nonsupplemented cystorraphies in specimens from canine cadavers.

Expressions of matrix metalloproteinases-1 and -9 and opioid growth factor in rabbit cornea after lamellar keratectomy and treatment with 1% nalbuphine.

PURPOSES: To evaluate the effects of nalbuphine 1% on the expression of metalloproteinase 1 (MMP-1), metalloproteinase 9 (MMP-9), and opioid growth factor (OGF) in rabbit corneas after lamellar keratectomy. METHODS: The rabbits were assigned to two groups: group nalbuphine (GN, n=30), which received 30 µL of nalbuphine 1% in 4 daily applications at regular intervals until corneal epithelialization, and group control (GC, n=30), which received physiological saline solution under the same conditions adopted in GN. The corneas were collected for immunohistochemistry on days 1, 3, 5, 7, and 9 after lamellar keratectomy, and the expressions of MMP-1, MMP-9, and OGF were analyzed. RESULTS: The expressions of MMP-1 and MMP-9 increased until day 5 of the evaluation, with no differences observed between GN and GC (p>0.05). On days 7 and 9, significant reductions were observed in the expression of MMP-1 (p<0.01), with no differences observed between GN and GC (p>0.05). The expression of OGF was constant in all periods (p>0.05), restricted to the corneal epithelium, and there was no difference between the groups (p>0.05). CONCLUSIONS: The study results showed that nalbuphine 1% did not alter the expression patterns of MMP-1, MMP-9, and OGF in rabbit corneas after lamellar keratectomy.

Expressions of matrix metalloproteinases-1 and -9 and opioid growth factor in rabbit cornea after lamellar keratectomy and treatment with 1% nalbuphine / Metaloproteinases de matriz 1, 9, e fator de crescimento opióide, em córneas de coelhos depois de ceratotomia e tratamento com nalbufina a 1%

ABSTRACT Purposes: To evaluate the effects of nalbuphine 1% on the expression of metalloproteinase 1 (MMP-1), metalloproteinase 9 (MMP-9), and opioid growth factor (OGF) in rabbit corneas after lamellar keratectomy. Methods: The rabbits were assigned to two groups: group nalbuphine (GN, n=30), which received 30 µL of nalbuphine 1% in 4 daily applications at regular intervals until corneal epithelialization, and group control (GC, n=30), which received physiological saline solution under the same conditions adopted in GN. The corneas were collected for immunohistochemistry on days 1, 3, 5, 7, and 9 after lamellar keratectomy, and the expressions of MMP-1, MMP-9, and OGF were analyzed. Results: The expressions of MMP-1 and MMP-9 increased until day 5 of the evaluation, with no differences observed between GN and GC (p>0.05). On days 7 and 9, significant reductions were observed in the expression of MMP-1 (p<0.01), with no differences observed between GN and GC (p>0.05). The expression of OGF was constant in all periods (p>0.05), restricted to the corneal epithelium, and there was no difference between the groups (p>0.05). Conclusions: The study results showed that nalbuphine 1% did not alter the expression patterns of MMP-1, MMP-9, and OGF in rabbit corneas after lamellar keratectomy. .

RESUMO Objetivos: Avaliar os efeitos da nalbufina 1% sobre a expressão da metaloproteinase 1 (MMP-1), da metaloproteinase 9 (MMP-9) e do fator de crescimento opióide (OGF), em córneas de coelhos submetidas à ceratectomia lamelar. Métodos: Constituíram-se dois grupos: grupo nalbufina (GN, n=30), que recebeu 30 µL de nalbufina 1% em 4 aplicações diárias, a intervalos regulares, até a epitelização corneal; controle (GC, n=30), que recebeu solução salina nas mesmas condições adotadas no GN. As córneas foram colhidas para imuno-histoquímica decorridos 1, 3, 5, 7 e 9 dias das ceratectomias lamelares, visando a se avaliarem as MMP-1, MMP-9 e OGF. Resultados: A expressão das MMP-1 e de MMP-9 se elevou até o quinto dia de avaliação, sem diferença entre GN e GC (p>0,05). Nos dias 7 e 9, observou-se redução significativa na expressão das enzimas (p<0,01), sendo que diferenças não foram observadas entre os grupos (p>0,05). O OGF exibiu imunomarcação constante em todos os períodos (p>0,05), restrita ao epitélio corneal. Não foram encontradas diferenças entre os grupos (p>0,05). Conclusões: Com base dos resultados obtidos, há como admitir que a nalbufina 1% não alterou o padrão de expressão da MMP-1, da MMP-9 e do OGF em córneas de coelhos submetidas à ceratectomia lamelar. .

Ketorolac eye drops reduce inflammation and delay re-epithelization in response to corneal alkali burn in rabbits, without affecting iNOS or MMP-9.

PURPOSES: To assess the effects of 0.5% ketorolac tromethamine without preservatives on the expression of iNOS and MMP-9 in alkali burn ulcers. METHODS: Twelve eyes of 120-day-old male rabbits were treated (TG) every 6 h with 0.5% ketorolac tromethamine and 12 other eyes were treated with saline solution (CG), immediately after the occurrence of ulcers by 1 M sodium hydroxide (NaOH). Re-epithelialization was monitored using fluorescein every 6 h. After 24 h, six corneas (n=6) of each group were collected (M1). The others (n=6) were collected after reepithelialization (M2). At both moments, the inflammatory infiltrate and the conditions of the newly formed epithelium were histologically analyzed. iNOS and MMP-9 were evaluated by immunohistochemistry. RESULTS: Mean epithelialization time in TG was 55 ± 0.84 h. In CG, it was 44 ± 1.06 h (p=0.001). At M1, corneas of TG had lower inflammatory exudation compared with (p <0.001). At M2, TG revealed discrete inflammatory exudation (p>0.05) and lower numbers of epithelial layers compared with CG. The mean iNOS in stromal cells did not differ in TG over both moments compared with CG (p>0.05) At M2, the central corneal region expressed more iNOS in both groups compared with the peripheral region. No significant differences were observed in iNOS scores of epithelial immunostaining between the groups and across M1 and M2 (p=0.69). Epithelial immunostaining scores for MMP-9 did not differ in TG compared with CG (p=0.69). The average immunostaining score of MMP-9 in stromal cells showed no differences between groups or moments. There was no correlation between immunostaining of iNOS and MMP-9 or between the amount of inflammatory cells and immunostaining of iNOS. CONCLUSIONS: Use of 0.5% keratolac tromethamine reduced inflammation and delayed reepithelialization in a cornea alkali burn model without impacting the expression of iNOS or MMP-9.

Effects of tramadol on tear production, intraocular pressure, and pupil size in dogs: clinical study / Efeitos do tramadol sobre a produção lacrimal, a pressão intraocular e o diâmetro pupilar de cães: estudo clínico

This study aimed to evaluate the effects of tramadol on tear production, intraocular pressure (IOP) and pupil diameter (PD) in healthy dogs. Dogs were randomly assigned to receive 4mg kg-1 (n=11) and 6mg kg-1 (n=11) of tramadol hydrochloride intramuscularly. Tear production (Schirmer tear test, STT-1), IOP (applanation tonometry) and the PD (electronic pachymetry) were assessed before, 30 and 60 minutes after administration of tramadol. Data were compared by analysis of variance for repeated measures (P<0.05). Parameters evaluated before, at 30 and 60min, in dogs treated with 4 and 6mg kg-1, were respectively: (STT-1) 22.50±3.38, 21.14±3.94 and 21.09±2.99mm min-1; and 23.05±3.73,22.64±3.76 and 22.82±3.25mm min-1. (IOP) 18.14±2.68, 17.68±2.59 and 18.23±3.84mmHg; and 19.05±2.27, 18.91±2.74 and 17.64±2.34mmHg. (PD) 6.71±0.65, 7.22±1.42 and 6.90±1.39mm; and 6.25±1.08, 6.80±1.27 and 6.49±0.90mm. All parameters evaluated did not change significantly among time points and dose regimen. Based on the conditions under which the experiments were conducted, tramadol did not affect tear production, IOP and PD in dogs, and could be used as a preoperative analgesic for intraocular surgery and pain control for any cause in patients affected by uveitis, glaucoma and keratoconjunctivitis sicca.

Objetivou-se estudar os efeitos da administração sistêmica do tramadol sobre a produção lacrimal, a pressão intraocular (PIO) e o diâmetro pupilar (DP) em cães saudáveis. Os cães foram aleatoriamente tratados com 4mg kg-1 (n=11) e 6mg kg-1 (n=11) de cloridrato de tramadol por via intramuscular. A produção lacrimal (Teste da lágrima de Schirmer, TLS-1), a PIO (tonometria de aplanação) e o DP (paquimetria eletrônica) foram mensurados antes, 30 e 60min após a aplicação do tramadol. Os dados obtidos foram avaliados pelo teste de análise de variância para medidas repetidas (P<0,05). Os parâmetros avaliados antes, aos 30 e aos 60min, em cães tratados, respectivamente, com 4 e 6mg kg-1, foram de: (TLS-1) 22,50±3,38, 21,14±3,94 e 21,09±2,99mm min-1; e 23,05±3,73, 22,64±3,76 e 22,82±3,25mm min-1. (PIO) 18,14±2,68, 17,68±2,59 e 18,23±3,84mmHg; e 19,05±2,27, 18,91±2,74 e 17,64±2,34mmHg. (DP) 6,71±0,65, 7,22±1,42 e 6,90±1,39mm; e 6,25±1,08, 6,80±1,27 6,49±0,90mm. Não houve diferença significativa entre os tempos e doses estudadas para quaisquer variáveis. Nas condições deste estudo, o tramadol não alterou a produção lacrimal, a PIO e o DP de cães, podendo ser utilizado como analgésico pré-operatório em cirurgias intraoculares e no controle da dor oriunda de qualquer etiologia em pacientes acometidos por uveíte, glaucoma e ceratoconjuntivite seca.

Ketorolac eye drops reduce inflammation and delay re-epithelization in response to corneal alkali burn in rabbits, without affecting iNOS or MMP-9 / Colírio de cetorolaco reduz inflamação e atrasa a epitelização em resposta a queimadura por álcali em coelhos, sem afetar iNOS ou MMP-9

Purposes: To assess the effects of 0.5% ketorolac tromethamine without preservatives on the expression of iNOS and MMP-9 in alkali burn ulcers. Methods: Twelve eyes of 120-day-old male rabbits were treated (TG) every 6 h with 0.5% ketorolac tromethamine and 12 other eyes were treated with saline solution (CG), immediately after the occurrence of ulcers by 1 M sodium hydroxide (NaOH). Re-epithelialization was monitored using fluorescein every 6 h. After 24 h, six corneas (n=6) of each group were collected (M1). The others (n=6) were collected after reepithelialization (M2). At both moments, the inflammatory infiltrate and the conditions of the newly formed epithelium were histologically analyzed. iNOS and MMP-9 were evaluated by immunohistochemistry. Results: Mean epithelialization time in TG was 55 ± 0.84 h. In CG, it was 44 ± 1.06 h (p=0.001). At M1, corneas of TG had lower inflammatory exudation compared with (p <0.001). At M2, TG revealed discrete inflammatory exudation (p>0.05) and lower numbers of epithelial layers compared with CG. The mean iNOS in stromal cells did not differ in TG over both moments compared with CG (p>0.05) At M2, the central corneal region expressed more iNOS in both groups compared with the peripheral region. No significant differences were observed in iNOS scores of epithelial immunostaining between the groups and across M1 and M2 (p=0.69). Epithelial immunostaining scores for MMP-9 did not differ in TG compared with CG (p=0.69). The average immunostaining score of MMP-9 in stromal cells showed no differences between groups or moments. There was no correlation between immunostaining of iNOS and MMP-9 or between the amount of inflammatory cells and immunostaining of iNOS. Conclusions: Use of 0.5% keratolac tromethamine reduced inflammation and delayed reepithelialization in a cornea alkali burn model without impacting the expression of iNOS or MMP-9. .

Objetivos: Avaliarem-se os efeitos do cetorolaco de trometamina 0,5%, sem conservante, sobre a expressão da iNOS e da MMP-9, em córneas com úlceras químicas. Métodos: Doze olhos de coelhos machos, 120 dias de idade, foram tratados (GT ), a cada 6 horas, com o cetorolaco de trometamina 0,5% e outros 12 com solução salina (GC), imediatamente à ocorrência de úlceras por hidróxido de sódio (NaOH) 1 mol/L. A reepitelização foi monitorada por fluresceína a cada seis horas. Decorridas 24 horas, seis córneas (n=6) de cada grupo foram colhidas (primeiro momento). As demais (n=6) o foram após a sua reepitelização (segundo momento). Em ambos os momentos, avaliaram-se o infiltrado inflamatório e as condições do epitélio neoformado (HE). Por imuno-histoquímica, avaliou-se a imunomarcação de iNOS e de MMP-9. Resultados: A média do tempo de epitelização no GT foi de 55 ± 0,84 horas. No GC, ela foi de 44 ± 1,06 horas (p=0,001). Às 24 horas, as córneas do GT apresentaram menor exsudação inflamatória (p<0,01). No segundo momento, o GT mostrou discreta exsudação inflamatória (p>0,05) e menor número de camadas epiteliais comparativamente ao GC. A média de imunomarcação de iNOS em células do estroma não diferiu do GT, em ambos os momentos (p>0,05). No segundo momento, a região central da córnea expressou mais iNOS, comparativamente à periférica, em ambos os grupos. Não se observaram diferenças significativas nos escores de imunomarcação epitelial de iNOS entre os grupos e os momentos (p=0,69). Os escores de imunomarcação epitelial para MMP-9 não diferiram entre os grupos (p=0,69). A média de imunomarcação da MMP-9 em células do estroma não exibiram diferenças entre os grupos e momentos da avaliação (p=0,32). Não houve correlação entre a imunomarcação de iNOS e de MMP-9, assim como quanto ao quantitativo de células inflamatórias e à imunomarcação de iNOS. Conclusões: Cetorolaco 0,5% reduziu a inflamação e atrasou a epitelização na queimadura corneal ...

Semen parameters and seminal plasma protein and biochemical profiles of dogs with benign prostatic hyperplasia after botulinum toxin type A intraprostatic injection / Parâmetros seminais e perfis bioquímicos e proteicos do plasma seminal de cães com hyperplasia prostática benigna após a administração intra-prostática de toxina botulínica tipo A

This study aimed to determine the effects of different concentrations of botulinum toxin type A (BT-A) on semen parameters, and seminal plasma biochemical and protein profiles of dogs with benign prostatic hyperplasia (BPH). Eighteen sexually intact male dogs with BPH were randomly divided in three groups, and received an intraprostatic injection of saline solution (control group - CG), 250UI (GI) or 500UI (GII) of BT-A under transabdominal ultrasound guidance. Semen was collected at baseline, 2, 4 and 8 weeks after treatment. Semen parameters were determined and seminal plasma pH, total protein (TP), total chlorides (TC), calcium (Ca), potassium (K), and sodium (Na) concentrations were assessed. One-dimensional sodium dodecyl sulfatepolyacrilamide gel eletrophoresis (SDS- PAGE) was performed to determine seminal plasma protein profile. Sperm parameters and seminal plasma pH, TP, TC, Ca and K mean values did not change significantly at any time point and among treated groups (P>0.05). The SDS-PAGE analysis of the pooled fractions identified 31 protein bands with molecular weights ranging from 3.9 to 106.2kDA in all treatment groups during the entire evaluation period. Regardless the used dose, intraprostatic BT-A injection do not alter semen parameters and seminal plasma biochemical and protein profiles of dogs with BPH.

O objetivo do presente estudo foi determinar a ação de diferentes concentrações de toxina botulínica tipo A (TB-A) sobre os parâmetros seminais, perfis bioquímicos e proteicos do plasma seminal de cães com hiperplasia prostática benigna (HPB). Dezoito cães hígidos, não orquiectomizados com HPB foram divididos em três grupos, os quais foram submetidos à injeção intra-prostática de solução salina (grupo controle - GC), 250UI (GI) ou 500UI (GII) de TB-A. Amostras seminais foram coletadas previamente aos tratamentos e após 2, 4 e 8 semanas. Os parâmetros seminais assim como os valores de pH e concentrações de proteínas totais (TP), cloretos totais (CT), cálcio (Ca), potássio (K), sódio (Na) do plasma seminal foram mensurados após as coletas. O perfil proteico do fluido prostático foi estabelecido por meio de eletroforese SDS-PAGE. Não foram constatadas diferenças significativas quanto aos parâmetros espermáticos e perfil bioquímico do plasma seminal intragrupos e intergrupos (P>0,05). À SDS-PAGE foram identificadas 31 bandas proteicas com pesos moleculares de 3,9 a 106,2kDA, em todos os tratamentos e durante todo o período de avaliação. Dessa forma, concluiu-se que, independentemente da dose utilizada, a injeção intra-prostática de TB-A não altera os parâmetros seminais, assim como os perfis bioquímico e proteico do plasma seminal de cães com HPB.